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Modulation of Gene Expression Using Zinc Finger-Based Artificial Transcription Factors

  • Sabine Stolzenburg
  • Alan Bilsland
  • W. Nicol Keith
  • Marianne G. Rots
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 649)

Abstract

Artificial transcription factors (ATFs) consist of a transcriptional effector domain fused to a DNA-binding domain such as an engineered zinc finger protein (ZFP). Depending on the effector domain, ATFs can up- or downregulate gene expression and thus represent powerful tools in biomedical research and allow novel approaches in clinical practice. Here, we describe the construction of ATFs directed against the promoter of the epithelial cell adhesion molecule and against the promoter of the RNA component of telomerase. Methods to assess DNA binding of the engineered ZFP as well as to determine and improve the cellular effect of ATFs on (endogenous) promoter activity are described.

Key words

Transient gene expression modulation VP64 Krüppel-associated box (KRAB) artificial transcription factors valproic acid 

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Sabine Stolzenburg
    • 1
  • Alan Bilsland
    • 2
  • W. Nicol Keith
    • 2
  • Marianne G. Rots
    • 1
  1. 1.Department of Pathology and Medical BiologyUniversity Medical Center GroningenGroningenThe Netherlands
  2. 2.Cancer Research UK Beatson LaboratoriesCentre for Oncology and Applied Pharmacology, University of GlasgowGlasgowUK

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